agar and Necrosis

High intensity ultrasound has shown considerable ability to produce precise and deep thermal coagulation necrosis. Focused, cylindrical, spherical or plane transducers have been used to induce high temperatures in tissues to coagulate proteins and kill cells. Recently magnetic resonance imaging (MRI) has been used, with extracorporeal or intracavitary focused transducers and cylindrical interstitial applicators, to monitor temperature distribution and provide feedback during heating procedures. If intraluminal applicators are used, the active part is in contact with the region of interest and it is essential to provide an accurate view of heat deposition and the extent of coagulation necrosis close to the transducer. The purpose of this study was to develop a 10 mm diameter intraluminal ultrasound applicator, designed to treat oesophageal cancers and compatible with MRI "real-time" temperature mapping. The active part of the ultrasound applicator, covered by a latex balloon, is a 15 X 8 mm2 plane transducer, which is in contact with the tumours during treatment. Each ultrasound exposure generates coagulation necrosis, in an area with the approximate shape of a rectangular parallelepiped up to 10 mm deep. When the exposures were repeated by rotating the applicator on its axis, sector-based or cylindrical volumes of necrosis could be produced, matching the shape of oesophageal cancers. Ex vivo trials were performed to demonstrate the applicator's compatibility with a clinical MRI scanner (1.5 T). MRI signals were acquired without any magnetic susceptibility distortion, even close to the applicator. Fast (0.72 images per second) 2D temperature mapping was performed during ultrasound exposure, using temperature-related proton resonance frequency shift at a resolution of 0.5 degrees C. Coagulation necrosis viewed with inversion recovery sequences, were in good agreement with the qualitative macroscopic observations made for the few cases tested in this study.

Topics: Agar; Animals; Esophageal Neoplasms; Liver; Magnetic Resonance Imaging; Necrosis; Swine; Temperature; Time Factors; Ultrasonic Therapy

The microbiota of root caries lesions of different grades of severity were studied. Fourteen lesions were examined. The experimental design of the study allowed correlation of histopathologically distinguishable stages with specific and distinct microbial populations. Dentin samples were ground in a sterile mortar and cultured anaerobically on nonselective Columbia blood agar plates supplemented with 5% hemolyzed human blood and on media selective for Lactobacillus spp. and streptococci. The cultivable microbiota were quantitatively speciated using Rapid ID 32A, Rapid ID 32 Strep, API 20 Strep, APIZYM, and API50 CH tests and SDS-PAG electrophoresis. In initial as well as in advanced lesions gram-positive bacteria accounted for approximately 90% of the CFUt. The proportion of Actinomyces, and in particular A. naeslundii was significantly higher (p < 0.05) in initial lesions than in advanced lesions. In contrast, the percentage of Streptococcus and especially S. mutans was higher (p < 0.05) in advanced than in initial lesions. Surprisingly low (0.8% of the CFUt) was the percentage of lactobacilli in advanced lesions. Gram-negative bacteria formed a minor part of the microbiota in both initial and advanced lesions. Among the gram-negative isolates, Prevotella, Selenomonas, and Bacteroides spp. were most noticeable. In advanced lesions, only the outermost layer of 0.5 mm thickness was populated by a high number of bacteria; the following segments harbored a negligible number of bacteria only. It is concluded that root caries is a continuous destruction process which is restricted to a subsurface zone of limited depth. The necrotic dentin is successively worn away, leading to a saucer-shaped cavitation which is repopulated by plaque. The creation of cavitations favors an aciduric flora. This might explain the succession of bacterial populations observed during the destruction process.

Topics: Actinomyces; Adult; Agar; Aged; Bacteria; Bacteroidaceae; Bacteroides; Blood; Colony Count, Microbial; Culture Media; Dental Plaque; Dentin; Disease Progression; Electrophoresis, Polyacrylamide Gel; Gram-Positive Bacteria; Humans; Lactobacillus; Middle Aged; Necrosis; Prevotella; Root Caries; Sodium Dodecyl Sulfate; Streptococcus; Streptococcus mutans

An investigation was made of the cell populations that occur in the sequential strata of human melanoma ( MM96 ) colonies. Colonies were either grown for the full duration of culture in a 'physiological' atmosphere of 5% O2 (unperturbed colonies), or grown in this atmosphere followed by a final incubation in a hypoxic atmosphere of less than 0.1% O2. Both autoradiographic and ultrastructural studies indicated that cell changes similar to those which occur successively in monolayer cultures experiencing nutritional deficiency, exist concurrently in the sequential strata of the larger unperturbed colonies. At the margin with the necrotic core, approximately half of the cells showed morphological changes associated with death by apoptosis. The other half were undergoing necrosis. Observations on colonies incubated for the final 24 or 48 h in a hypoxic (less than 0.1% O2) atmosphere showed that many of these cells, although otherwise well preserved, developed oedema complicated by cytoskeletal rupture and extrusion of areas of damaged cytoplasm within membrane-bound vesicles. Although sudden-onset hypoxia did not appear to precipitate cell death in small colonies previously lacking a necrotic core, large colonies suffered a marked reduction in the width of their viable rims. Cell death under this circumstances was by necrosis, the same mode of death as occurs with infarction. The study indicated that apoptosis was associated with sub-acute cell death as occurs with progressive nutrient depletion and catabolite accumulation, whereas necrosis was associated with acute cell death as seen in previously compromised cells subject in addition to sudden-onset hypoxia.

Topics: Agar; Aged; Cell Line; Cell Survival; Female; Humans; Kinetics; Melanoma; Microscopy, Electron; Necrosis; Oxygen; Tumor Stem Cell Assay

Chinese hamster ovary (CHO) cells plated on agar form two classes of colonies; those which increase continuously in diameter and those which become arrested in outward growth. All colonies continue to increase in mass and thickness as demonstrated by computer-assisted analysis of time sequence photographs of several thousand colonies and by examination of histological sections. Colonies which shift from a monolayer to a mounded morphology at fairly large colony diameter (greater than or approximately 1 mm) continue to increase in diameter. Colonies in which mounding occurs at smaller colony diameter (less than or approximately 1 mm) cease to increase in diameter but continue to increase in thickness, as demonstrated by histological examination and by computer-assisted analysis. Rapid cell division occurs at the edge of all colony classes, as shown by the distribution of mitotic figures. In arrested colonies these dividing cells must move toward the colony core to compensate for dying cells. Necrotic cells are found as a discrete zone at the air-colony interface in all cases for CHO cell colonies growing on agar. As such, necrosis is probably due to limitations in nutrient diffusion upward from the agar rather than oxygen diffusion downward.

Topics: Agar; Animals; Cell Division; Cell Line; Cell Survival; Clone Cells; Computers; Cricetinae; Kinetics; Mitotic Index; Necrosis

Topics: Agar; Breast Neoplasms; Cell Differentiation; Electrophoresis; Embryo, Mammalian; Female; Humans; Isoenzymes; L-Lactate Dehydrogenase; Mitosis; Necrosis

Topics: Agar; Animals; Carbamates; Endotoxins; Epinephrine; Escherichia coli; Hemorrhage; Necrosis; Pyridines; Rabbits; Thrombosis

Topics: Acetates; Acute Disease; Agar; Animals; Bile; Calcium Chloride; Carrageenan; Chlorides; Copper; Croton Oil; Ethanol; Female; Formaldehyde; Glucose; Histamine; Indium; Lead; Manganese; Mercury; Necrosis; Papain; Phosphates; Polymyxins; Polysaccharides; Rats; Skin Diseases; Tannins; Trypsin; Zinc

Topics: Agar; Animals; Common Bile Duct; Duodenal Diseases; Female; Hemorrhage; Jejunum; Necrosis; Rats; Thrombosis